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OfflineMr.Christmas
Who?
I'm a teapot


Registered: 04/08/20
Posts: 119
Loc: USA
Last seen: 9 days, 45 minutes
Contam Central after switching to SAB
    #26864903 - 08/05/20 04:01 PM (3 years, 6 months ago)

So, I know that everyone here is a big hater of glove boxes but ever since I switched to SAB, I haven't had a single clean Agar plate. My sanitation procedures haven't changed, the spores I use haven't changed, the agar I use hasn't changed, but hot diggity has the contame been steady.

Has anyone else experienced this? Should I do some modifications to the SAB to make it better? I'm honestly considering my just buying a flow hood now.


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Todo lo que hagas en la vida será insignificante; pero es muy importante que lo hagas.


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OfflineTipOfTheCap
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Registered: 07/31/20
Posts: 73
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Re: Contam Central after switching to SAB [Re: Mr.Christmas]
    #26865000 - 08/05/20 04:49 PM (3 years, 6 months ago)

I'm sort of green, but:

I use my SAB upside down. I spray the inside with 70 percent iso and wipe down. I iso my table and wipe. I then take a fresh towel and lay it down on the table. I spray the towel with soap water heavily and I spray the inside of the SAB with soap water. I put my materials in, and turn off my AC and close the room door. I wait 3 minutes for the air to settle in the room and inside the SAB before wiping my arms and gloves down with iso before proceeding. The idea is that particles will fall to the towel and get stuck in the soap and fibers. And the soap is sticky everywhere inside the SAB walls. I've had great results. Better than using a lid.

Are you using powdered gloves by chance? That got me once.

Sound like your materials are pretty sterile and the only variable was the glove box change. As simple as it seems, small details can make the difference. Is your PC acting okay?


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Invisibledfwerydfhg
Stranger
Registered: 05/04/20
Posts: 194
Re: Contam Central after switching to SAB [Re: TipOfTheCap]
    #26865135 - 08/05/20 05:31 PM (3 years, 6 months ago)

Sterile technique (where and how you move your hands, arms, and instruments, not what kind of stuff you spray) is going to be very different between the two systems. How much have you changed that?


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OfflineMarmie
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Registered: 10/21/15
Posts: 918
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Re: Contam Central after switching to SAB [Re: dfwerydfhg]
    #26865156 - 08/05/20 05:41 PM (3 years, 6 months ago)

If the holes are to small it can cause a piston effect, try some incence in there n see if your getting a current, without doing work ofcourse jst a sort of test run


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OfflineMr.Christmas
Who?
I'm a teapot


Registered: 04/08/20
Posts: 119
Loc: USA
Last seen: 9 days, 45 minutes
Re: Contam Central after switching to SAB [Re: Marmie]
    #26865657 - 08/05/20 11:35 PM (3 years, 6 months ago)

Quote:

TipOfTheCap said:
I'm sort of green, but:

I use my SAB upside down. I spray the inside with 70 percent iso and wipe down. I iso my table and wipe. I then take a fresh towel and lay it down on the table. I spray the towel with soap water heavily and I spray the inside of the SAB with soap water. I put my materials in, and turn off my AC and close the room door. I wait 3 minutes for the air to settle in the room and inside the SAB before wiping my arms and gloves down with iso before proceeding. The idea is that particles will fall to the towel and get stuck in the soap and fibers. And the soap is sticky everywhere inside the SAB walls. I've had great results. Better than using a lid.

Are you using powdered gloves by chance? That got me once.

Sound like your materials are pretty sterile and the only variable was the glove box change. As simple as it seems, small details can make the difference. Is your PC acting okay?




Non-powdered gloves. I'm going to try turning off my AC, although this usually hasn't caused an issue.

Quote:

dfwerydfhg said:
Sterile technique (where and how you move your hands, arms, and instruments, not what kind of stuff you spray) is going to be very different between the two systems. How much have you changed that?




Hmm, not much if I'm being honest, what do you mean by how I move my arms? I've seen videos or people demonstrating their SAB and it doesn't seem any different from what I'm doing.

Quote:

Marmie said:
If the holes are to small it can cause a piston effect, try some incence in there n see if your getting a current, without doing work ofcourse jst a sort of test run




This is a really fantastic idea. I'll definitely try that this weekend.


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Todo lo que hagas en la vida será insignificante; pero es muy importante que lo hagas.


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OfflineMarmie
Peter peter portion eater
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Registered: 10/21/15
Posts: 918
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Last seen: 14 days, 12 hours
Re: Contam Central after switching to SAB [Re: Mr.Christmas]
    #26865670 - 08/05/20 11:54 PM (3 years, 6 months ago)

If your running an ac while doing it defanitly turn that off,  in a spawn room its fine to have em on but when doing clean work its best to have them off, if thats on while your working thats probably your issue,  keep your furnice and ac’s off while workin it’s unnecessary air movement and most are usualy pretty dirty.

And by your arms he means slow movements , making sure your hands never go overtop recieving/donor jars or plates.


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Offlinerocklobster
Walking my friends home...

Registered: 07/10/20
Posts: 50
Loc: Savage Reservation
Last seen: 3 months, 7 hours
Re: Contam Central after switching to SAB [Re: Marmie]
    #26882526 - 08/15/20 11:46 PM (3 years, 5 months ago)

I've had great success so far using Bod's Easy AF SAB TEK

I've followed it exactly even down to the same 110qt tub. My arm holes were cut with a 5" hole saw so they're a little smaller than BOD's. I use a 2x4 foot plastic table (Lifetime brand from Home Depot). No lid, table surface as the floor and no towel or racks. AC off, no fans on.

Once I have the SAB and table sprayed with soapy water, I add my plates by cutting the bottom of the bag, carefully exposing the bottom of the bottom plate and setting it on the edge of the table. I slowly remove the bag without lifting the lids of any of the plates. I then gently lift the edge of the box and slowly slide them in, trying to minimize air movement. I try to make all of my movements deliberately, but honestly my first time using it I was a mess. I still had no contam on my control plates. 

I'm PCing my media for 25 mins using the standard 10/10 per 500ml MEA recipe. I use the slimline 500ml Pyrex bottles, in a 23qt Presto. I pour when my media bottle reaches 117f (again per one of BOD's teks). I let it cool inside of the SAB and open the lid for the first time in the box. Lid is placed on the floor and it's not put back on until the bottle is empty.

I take a shower before pouring and spray my gloves and arms down with ISO before I begin. I pour all in one sitting without getting up to minimize pulling my arms in and out.

I wrap my plates as soon as they're cooled (5mins) using Gladwrap cut in two inch rolls. I can leave them for a few weeks at room temp before pouring at that point.

I take no credit for my methods here, I've read nearly every one of the trusted TEKs cover to cover and try to follow them to a T. If you hit each of the points I'm confident you'll have success as well.


Edited by rocklobster (08/15/20 11:47 PM)


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OfflineYour Dudeness
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Registered: 11/10/18
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Re: Contam Central after switching to SAB [Re: rocklobster]
    #26882582 - 08/16/20 12:40 AM (3 years, 5 months ago)

Agar without a flow hood you can't mess around. 

Putting your arms in without clothing is contam reason #1.  Try wearing a long sleeved shirt.  Use a respirator too.  RR says aluminum foil is sterile from the factory so tear off a couple sheets and use that as your work surface.


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RR sent me


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OfflineA.k.aM
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Registered: 10/27/19
Posts: 16,816
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Re: Contam Central after switching to SAB [Re: Your Dudeness]
    #26882833 - 08/16/20 06:32 AM (3 years, 5 months ago)

I can’t imagine going from glovebox to SAB causing huge problems.

The only difference would be if your bare forearms are now going in there. Try wiping them down with iso first maybe.

Otherwise it’s gotta be the technique cuz it’s basically the same thing.


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LAGM2020


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OfflineChapuco
nice cock bro, nice tits miss
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Registered: 08/11/20
Posts: 30
Last seen: 2 years, 8 months
Re: Contam Central after switching to SAB [Re: Mr.Christmas]
    #26882858 - 08/16/20 07:01 AM (3 years, 5 months ago)

check your arms, cover them with film, skin particles could be contaminating your dishes. Also use gloves and spray them with iso alcohol 70%


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Offlinejcm4620
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Registered: 05/26/19
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Re: Contam Central after switching to SAB [Re: Chapuco]
    #26882875 - 08/16/20 07:13 AM (3 years, 5 months ago)

way more of us use sab instead of a fh and most have no issue at all. id rather have a fh but a sab can be every bit as effective.  so ya is also wear clean long sleeve shirt with gloves i dont use a mask personally but it aint gnna hurt. a mask while workin with a sab is waaaay more effective than a mask at the store for covid but neither are 100% needed


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PANAEOLUS FRUITING MADE SIMPLE



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